Purification of G1 daughter cells from different Saccharomycetes species through an optimized centrifugal elutriation procedure.

Martial Marbouty (1) , Caroline Ermont (1) , Bernard Dujon (2) , Guy-Franck Richard (2) , Romain Koszul (1)

1 Régulation spatiale des Génomes - Spatial Regulation of Genomes
2 Génétique Moléculaire des Levures

Centrifugal elutriation discriminates cells according to their sedimentation coefficients, generating homogeneous samples well suited for genomic comparative approaches. It can, for instance, isolate G1 daughter cells from a Saccharomyces cerevisiae unsynchronized population, alleviating ageing and cell-cycle biases when conducting genome-wide/single-cell studies. The present report describes a straightforward and robust procedure to determine whether a cell population of virtually any yeast species can be efficiently elutriated, while offering solutions to optimize success. This approach was used to characterize elutriation parameters and S-phase progression of four yeast species (S. cerevisiae, Candida glabrata, Lachancea kluyveri and Pichia sorbitophila) and could theoretically be applied to any culture of single, individual cells.

Keywords: yeast; elutriation; replication; synchronization; Ascomyceta; daughter cells

Domains: Genetics

Martial Marbouty, Caroline Ermont, Bernard Dujon, Guy-Franck Richard, Romain Koszul. Purification of G1 daughter cells from different Saccharomycetes species through an optimized centrifugal elutriation procedure.. Yeast, 2014, 31 (5), pp.159-166. ⟨10.1002/yea.3005⟩. ⟨pasteur-01420008⟩